Casticin has wide-ranging functional activities, but its water solubility is poor in food products. Here, a nanoemulsion stabilized by Maillard whey protein isolate conjugates (MWPI) was fabricated to encapsulate casticin. The nanoemulsion, with an average diameter of 200 nm, possessed the capability to load 700 μg/g casticin. MWPI-stabilized nanoemulsion showed better stability than that of the WPI nanoemulsion during 4 weeks of storage. Both the inhibition effects of the casticin-loaded nanoemulsion on cancer cells and the process of cellular uptake were studied. Results revealed that the casticin-loaded nanoemulsion had better inhibitory activity in HepG2 and MCF-7 cells than free casticin. Cellular uptake of the nanoemulsion displayed a time-dependent manner. After the nanoemulsion passed into HepG2 and MCF-7 cells, it would locate in the lysosome but not in the nucleus. The main pathway for the nanoemulsion to enter HepG2 cells was pinocytosis, whereas, it entered MCF-7 predominantly through the clathrin-mediated pit. This work implies that MWPI-stabilized nanoemulsions could be utilized as an effective delivery system to load casticin and have the potential to be applied in the food and pharmaceutical industries. Vitex (Fructus Vititcis) [Casticin, 5,3′-dihydroxy-3,6,7,4-tetramthoxyflavone, is a kind of polymethoxy-flavonoid (PMF) extracted from the traditional Chinese medicine ititcis) . Numerouititcis) , anti-hyThe fabrication of an emulsion is a potential way to encapsulate the poorly water-soluble PMFs and improve their bioavailability. Because hydrophobic bioactive compounds are easy to dissolve in the oil phase, they can be kinetically stable dispersed into the aqueous phase after forming nanoemulsions. Due to the small size (20–200 nm), nanoemulsions have the advantages of a large specific surface area, high stability, and adjustable rheology properties ,8. When Glycosylate-modified protein, mainly through the Maillard reaction, possesses suitable amphiphilicity, which can provide steric hindrance to prevent the aggregation and disproportionation of emulsion droplets . MengíbaIn the current work, the Maillard whey protein isolate-lactose conjugate (MWPI) was prepared and utilized as an emulsifier to improve the stability of the nanoemulsion. Casticin was loaded in the MWPI-stabilized nanoemulsion to inhibit the growth of MCF-7 and HepG2 cancer cells. Finally, the kinetics and mechanism of the cellular uptake of nanoemulsion were preliminarily clarified.Vitex, and the purity was 94.6% after comparison with a standard by HPLC. The casticin standard , L-lactose, and dimethyl sulfoxide  were purchased from Aladdin Reagent Co., Ltd. . The MCF-7 and HepG2 cell lines were obtained from the China Center for Type Culture Collection . Fetal bovine serum (FBS) and Dulbecco’s modified eagle’s medium (DMEM) for cell culture were purchased from Gibco . Penicillin-streptomycin and 0.25% trypsin were obtained from Jinuo Biomedical Technology Co., Ltd. . Phosphate buffer saline (PBS) was purchased from Hyclone . Cell-culture flasks with a capacity of 25 cm2, 96-well cell-culture plates, and cryotubes were purchased from Corning Co., Ltd. . Nile red and coumarin-6 were purchased from Aladdin Reagent Co., Ltd. . Lyso-Tracker red  and 4,6-diamino-2-phenyl indole (DAPI) were purchased from Beyotime Biotechnology Co., Ltd.  and Solarbio Technology Co., Ltd. , respectively.The whey protein isolate (WPI) was gifted from Hilmar, USA. The casticin was extracted from w/w), and the final solid content was 5–10%. The mixture solution was adjusted to pH 7.0 and stirred for 12 h. After the solution was freeze-dried, the powders were placed in a desiccator with saturated KBr (relative humidity 79%). The WPI-lactose Maillard conjugates were obtained after reacting at 70 °C for 24 h. WPI-lactose Maillard conjugate was fabricated according to the method reported by Liu et al. with some modifications ,22. Brie−1. The resolution and the number of scans were 4 cm−1 and 32, respectively.The powders of WPI and WPI-lactose Maillard conjugates were mixed with KBr separately, and then they were pressed into thin slices. The FTIR spectra of powders were measured with a NEXUS670 spectrometer . The wavenumber ranged from 4000 to 400 cmThe sample (2 mg/mL) of WPI and MWPI were diluted 5 times with the loading buffer, centrifuged at 6000 rpm for 5 min, and then boiled for 5 min to complete denaturation. According to the protocol of the SDS-PAGE kit , the 12% separating gel and 5% concentrated gel were used for separation. Operation proceeded at 60 V for the first 30 min, and then changed to 120 V. Then, the gel was stained with 0.25% Coomassie Brilliant Blue R-250 for 30 min, and de-stained. An image was obtained using a Bio-Rad scanner .The WPI or MWPI powder was dissolved in 10 mM PBS (pH 7.4) at a concentration of 1.0 wt%, and the oil phase fraction was set at 10%. The MWPI-stabilized nanoemulsion was fabricated via high-speed homogenization  at 10,000 rpm for 10 min, and a high-pressure homogenization  with 900 Mbar and 10 cycles in succession.Nanoemulsions stabilized by WPI and MWPI were placed at 4 °C, and 0.02% sodium azide was added in emulsions to prevent bacteria growth. The particle size of nanoemulsions measured by Zetasizer Nano-ZS  at 1 week, 2 weeks, and 4 weeks was recorded as the indicator for evaluating storage stability. g for 15 min, 200 μL of the supernatant was mixed with 800 μL methanol and then filtered through a 0.45 μm filter for high performance liquid chromatography (HPLC) analysis. The samples (20 μL) were analyzed by a reverse-phase C18 column  with buffer A (ultrapure water) and buffer B (acetonitrile) at a flow rate of 1 mL/min. The detection wavelength was 254 nm. The gradient elution method was as follows: from 40% to 55% buffer B for 10 min, from 55% to 70% buffer B for 5 min, from 70% to 80% buffer B for 5 min, and held at 40% buffer B for 10 min.An excess of casticin was added to 10 g medium chain triglyceride (MCT), and magnetically stirred overnight at room temperature. After centrifugation at 5000× The CAS was dissolved in MCT in a saturated state as an oil phase. The water phase was a 1.0 wt% MWPI solution. The homogenization process was carried out as described in The MWPI-stabilized nanoemulsion was spray dried under the following conditions: inlet air temperature 115 °C, feed rate 14 mL/min, spray airflow 1000 L/h, and needle frequency 1 time/15 s. Finally, the obtained powder was kept in a sealed bag protected from light, at 4 °C.g for 10 min. Next, the content of casticin was analyzed using HPLC as described in The casticin in the nanoemulsion was extracted by adding 1 mL of n-hexane to 1 g of nanoemulsion, followed by vortexing and centrifugation at 10,000× The nanoemulsion was diluted 10-fold with DMEM  and stored at 37 °C for 24 h. The particle size and polydispersity (PDI) of the nanoemulsion were recorded using a Marlven Nanosizer ZS90 .MTT -2,5-diphenyltetrazolium bromide) was used to evaluate the ability of the nanoemulsion to inhibit the proliferation of MCF-7 and HepG2 cells. Briefly, cells were seeded on 96-well plates in the exponential growth phase, and then the nanoemulsion was diluted with DMEM to control the final concentrations of casticin. After incubation, the nanoemulsion was removed, and each well was washed with PBS 2~3 times. Finally, 100 μL DMEM and 10 μL MTT solution were added to the wells and incubated for 4 h. Then, the medium was removed and 150 μL DMSO/well was added to dissolve the formazan crystals. The absorbance at 570 nm was recorded on a microplate reader . Untreated cells were defined as the negative control, and their OD value was recorded as 100% survival.4 cells/mL) was inoculated into the CLSM dish and cultured for 24 h (37 °C and 5% CO2). After removing the medium and washing the cell surface with PBS, the casticin-loaded nanoemulsion labeled with Nile red was added and cultured for 10 min, 20 min, 30 min, 1 h, and 2 h. Before CLSM observation, the nanoemulsion was removed and the cells were washed three times by PBS buffer. The nuclei were stained with DAPI for 15 min.First, Nile red was added in the MCT as the fluorescence dye before preparing the nanoemulsion. Then, 1 mL of cell suspension .Each measurement of the samples was repeated at least three times, and the data was expressed as the average values and standard error in figures. The data were analyzed by SPSS 23  via one-way ANOVA. The significance level was −1 (N-H stretching and hydrogen bonding), 1640 cm−1 , 1540 cm−1 , and 1390 cm−1  [−1 was shifted to 3390 cm−1, which might be ascribed to the stretching vibration of free-OH or the deformation vibration of the N-H bond. The peak at 1640 cm−1 is derived from the α-helix or random coil structure. After modification, the peak of WPI at 1640 cm−1 was shifted to 1660 cm−1, indicating that the α-helix or random coil structure had been changed. Compared with WPI, MWPI had a new absorption peak at 1000–1260 cm−1, which might be attributed to the stretching vibration of the C-O bond, resulting from the presence of multiple hydroxyl groups on the sugar chain. The FTIR spectra of WPI and MWPI are shown in pansion) ,24. As fThe solubility of casticin in MCT was 2.05 mM. MCT is a medium-chain fatty acid with low viscosity, high transparency, and good compatibility and is widely applied in the pharmaceutical, cosmetic, and food industries. The casticin-saturated MCT and whey protein isolate-lactose conjugate were chosen to fabricate the nanoemulsion via high-speed homogenization and high-pressure homogenization, and then the nanoemulsion was spray-dried. According to HPLC analysis, the casticin content in the spray-dried casticin-loaded nanoemulsion powder was 700 μg/g. The stability of the casticin-loaded nanoemulsion in DMEM medium was characterized by evaluating the particle size. Because liver cancer and breast cancer are among the cancers with the highest incidence in China and in women in the world, respectively, the typical cell lines of Hep G2 and MCF-7 cell lines of the two were chosen as models. The inhibitory effect of casticin on HepG2 and MCF-7 and the cellular uptake mechanism might provide preliminary directions for cancer treatment. The anti-cancer activity of the blank emulsion, free casticin, and casticin-loaded nanoemulsion on MCF-7 and HepG2 cells was measured by the MTT test. The relationship between the dosage of casticin and cell viability is shown in The uptake of the casticin-loaded nanoemulsion in the MCF-7 and HepG2 cells observed by CLSM is shown in To clarify the mechanism of the inhibition of cancer cells by the casticin-loaded nanoemulsion, the determination of cell localization was carried out. The casticin-loaded nanoemulsion was labeled with coumarin-6 (green color); the cell nuclei were dyed with DAPI (blue color), and the lysosomes were labeled with LysoTracker red. As shown in Cellular uptake is the process by which cells absorb molecules or materials from the extracellular space. Nanoparticles are taken into cells mainly through endocytosis ,31, whicHerein, three pathway inhibitors were used to investigate some of the specific pathways involved during the cellular uptake of the casticin-loaded nanoemulsion. Dynasore inhibits dynein-dependent cellular uptake, such as clathrin-mediated and caveolae-mediated pits . ChlorprAs for HepG2 cells, the dynasore and amiloride HCl dihydrate inhibitors had more influence on the cellular uptake of the casticin-loaded nanoemulsion, but chlorpromazine showed limited inhibition. Therefore, macro-pinocytosis is likely the main pathway for the entry of the casticin-loaded nanoemulsion, as well as the involvement of a small amount of clathrin-mediated and caveolae-dependent pits. The results showed a little bit of similarity to silver nanoparticles  and siliTemperature affects the entry of nanoemulsions into cells. At a low temperature, the fluidity of the cell membrane become weak, and the activity of enzymes involved in uptake activities is significantly reduced, resulting in a decrease in energy-dependent cellular uptake. As shown in The nanoemulsion prepared by MWPI has better stability than that prepared by WPI. The casticin-loaded nanoemulsion was fabricated by a MWPI stabilized with MCT. The loading amount of casticin in the nanoemulsion after spray-drying reached 700 μg/g. The casticin-loaded nanoemulsion had good physical stability in the DMEM medium as its particle size was maintained at 200 nm over 8 h with a small PDI (<0.2). Compared with free casticin, the casticin-loaded nanoemulsion showed a stronger inhibitory effect on MCF-7 and HepG2 cells. The inhibitory effect was dependent on the dosage of casticin and the co-culturing time. The phagocytosis of the casticin-loaded nanoemulsion by MCF-7 and HepG2 cells was time-dependent. Over time, the cellular uptake of the casticin-loaded nanoemulsion increased. The casticin-loaded nanoemulsion entered the cell by co-localization with the intracellular lysosome, indicating that a portion of the casticin-loaded nanoemulsion entered the cell via the lysosome. The main mechanism for the casticin-loaded nanoemulsion to enter the cells was via pinocytosis for HepG2 and clathrin-mediated pits for MCF-7 cells.


Post-Market Clinical Follow-Up has been integrated into the new Medical Device Regulations since 2020. The CD Horizon Solera 4.75 mm instrumentation (CD-Solera) was introduced worldwide in 2009, and specifically intended for surgical treatment of pediatric and adolescent scoliosis patients. The objective of this study was to evaluate the safety and efficacy of the CD-solera 4.75 instrumentation in surgical treatment of adolescent idiopathic scoliosis (AIS).94 consecutive AIS patients, 82 female, 12 male, who underwent posterior correction with CD-Solera instrumentation between 2010 and 2016 at age 14.8 ± 1.6 years, were retrospectively included. The minimum follow-up was two years. On pre- and postoperative biplanar full spine radiographs Cobb angles of the primary and secondary curves and sagittal profile were measured before surgery, immediately postoperative, and at two-year follow-up. Medical records were reviewed for complications. Clinical outcome was analyzed using theSRS-22r questionnaire.In this study 77% of the patients had a structural thoracic curve (type Lenke 1 or 2), and 23% had a structural (thoraco-)lumbar curve (Lenke 3-6). A correction of 55.1% and 51.7% was achieved respectively immediately post-operative, and at last-year follow up for the primary curve. The mean loss of correction was 2°. Health related quality of life was 4.0 (good) on the SRS-22r-questionnaire. In total six revision operations were executed, of which one was related to the material (rod breakage). Other reasons for revision operation were not due to the material. No neurological problems were encountered.In patients with AIS the initial correction and maintenance of correction as achieved by posterior spinal fusion using the CD-Solera instrumentation, is comparable to other reported devices. Complication rates are low and health related quality of life comparable to literature. The CD-Solera can be regarded as a safe and effective instrumentation in surgical treatment of AIS. The assessment of quality in health care and in orthopedics has become more important in the last decade In scoliosis surgery, correction of the deformity is not solely related to the preoperative magnitude and rigidity of the curve, but also to the quality of the instrumentation used. Thus, the advantages and disadvantages of new instrumentation should be considered critically In 2017 Europe's new Medical Devices Regulations (MDR) were published. These regulations state that Post-Market Clinical Follow-Up will be integrated into the MDR to provide vital clinical evidence. Full implementation of the new MDR became a requirement in 2020 ,11. AmonThe CD Horizon Solera 4.75 mm Spinal System  (CD-Solera) was introduced worldwide in 2009. In our clinic, this instrumentation is used since 2010 for posterior operative correction of adolescent idiopathic scoliosis (AIS). The system consists of cobalt chrome multi-axial screws with dual lead thread form, two cobalt chromium extra strong (CoCr Plus) rods, standard laminar and pedicle hooks and modular crosslink plates. The presumed improvement of the CD-Solera system relative to its predecessors, the CD Horizon Legacy 4.5 and 5.5 systems, is a reduction in overall volume of the Multi-Axial Screws of 12% and 26%, respectively. The lower profile construct makes it more suitable for the younger and thinner patients. The dual lead thread form of the pedicle screws  enables enhanced fixation with reduced risk of screw pullout As yet, there is no published clinical data available about the CD-Solera instrumentation. With Europe's new MDR in mind, the purpose of this study is to determine the safety and efficacy of the CD-Solera instrumentation in the surgical treatment of AIS by means of a post-marketing surveillance with a minimum follow-up of two years.The used instrumentation in all patients was the CD Horizon Solera 4.75 mm Spinal System  (CD-Solera). The system consists of cobalt chrome multi-axial screws with dual lead thread form, two cobalt chromium extra strong 4.75mm (CoCr Plus) rods, standard laminar and pedicle hooks and modular crosslink plates.For this retrospective single center cohort study, we assessed clinical data from all consecutive patients with AIS who underwent surgical correction with the CD-Solera instrumentation from January 2010 until December 2016.Inclusion criteria were (1) age below 19 years at the time of surgery, (2) AIS (3) surgical correction with the CD-Solera instrumentation (4) standing full spine posterior-anterior and lateral radiographs pre-operative, immediately postoperative and at two-year follow-up. No patients who met the inclusion criteria were excluded or lost to follow-up. Threshold to operate in these AIS patients was a structural primary curve over 45°. This decision was made by the treating spine surgeon.We included 80 patients with AIS, 71 female, 9 male. The minimum length of follow-up was two years. Median length of follow up was 27 months (24-88 months).Demographic and surgical data were obtained from clinical records. These data included gender, age at surgery, body mass index (BMI), American Society of Anesthesiologists (ASA) classification, operated segments, number of fused spinal levels, implant density (we consider double fixation (left and right) per vertebra as an implant density of 100% An exploratory search in PubMed was performed to compare our results with the current literature.. Hybrid posterior instrumentation was performed with hooks cranially and pedicle screws caudally  with motor evoked potentials (MEP) and somatosensory evoked potentials (SSEP) monitoring ,15. Hybrally see . Curve cStanding full spine AP and sagittal radiographs were examined pre-operatively, post-operatively and at two-year follow-up. We measured the thoracic and thoracolumbar/lumbar curves and thoracic kyphosis (from superior endplate of T5 to the inferior endplate of T12) and lumbar lordosis (between lower endplate of T12 and upper endplate S1) using the Cobb method Radiographic measurements were carried out by a medical student (LS) and verified by an orthopedic spine surgeon (PW). These measurements were comparable within 3°, which is within the range of 0° to 5° what is commonly mentioned as an acceptable variation ,20. To d[21]Direct correction rate: (Pre-operative Cobb angle - Post-operative Cobb angle) / Pre-operative Cobb angle * 100 = … %Maintenance of correction: (Post-operative Cobb angle – Last follow-up Cobb angle) / Post-operative Cobb angle * 100 = … %Flexibility rate: (Pre-operative Cobb angle - Pre-operative side-bending Cobb angle) / Pre-operative Cobb angle * 100 = … %The Dutch Scoliosis Research Society-22r questionnaire (SRS-22r) was sent at minimum 2 years post-operative to the patients. Informed consent was obtained before sending out the SRS-22r questionnaires. The SRS-22r is a revised version of the older SRS-22 and its Dutch translation was earlier assessed as reliable and valid Statistical analysis was performed using IBM SPSS Statistics version 24. Descriptive statistics were used to determine the means, standard deviations (SD), and ranges. Means and SD were calculated for baseline characteristics, sagittal correction and SRS-22r. Ranges were calculated for the coronal correction. Prior to statistical analyses distribution of continuous variables were tested by skewness and kurtosis. P values were based on the paired samples T test for continuous variables. P-values equal to or below 0.05 were considered statistically significant.This study was approved by the Institutional Review Board of the Maastricht University Medical Centre, no. 2017-0253.Demographic and specific surgical data are listed in The mean number of fused levels was 10.5 ± 1.9 and the mean implant density was 76.4 ± 7.8%. Mean surgery time was 255 (± 64) minutes. Median blood loss was 400ml (100 – 1700 ml).The mean pre-operative primary angle was 58.7 ± 10.5⁰ and was corrected to 26.5 ± 10.6⁰ average, direct post-operative and to 28.5 ± 11.0⁰, at last follow-up. Mean pre-operative secondary angle was 39.2 ± 10.0⁰ and corrected to 21.8 ± 11.5⁰, direct post-operative and to 22.1 ± 12.2⁰, at last follow-up. A mean correction of 51.7% and 44.6% was achieved at last follow-up for the primary and secondary curve, respectively .Table 2PMean pre-operative kyphosis (T5-T12) was 22.2⁰ and reduced to 17.8, at last follow-up. Mean pre-operative lordosis was 54.5⁰ and 54.7⁰, at last follow-up .Table 3MIn In total nine complications were observed , for whiOne patient had a painful protruding hook which was covered by hardly any soft tissue, and therefore removed. In one patient the entire material was removed in two stages at 36 and 51 months follow-up because of invalidating pain and in one patient the total material was removed at 43 months follow-up due to mental health problems. This patient felt burdened with the back instrumentation and suffered from invalidating pain. She requested removal of the material.One revision surgery was performed because of ventral screw protrusion, which was fortuitously found at regular follow-up. The sixth revision surgery was performed because of deep wound infection two months post-operatively. Two patients had a superficial wound infection (2.1%) which resolved after three to four weeks of antibiotic treatment. No peri-operative or neurological complications were encountered.Apart from one case of rod breakage, none of the complications could be attributed to the CD-Solera instrumentation specifically.The current study was conducted to evaluate the safety and efficacy the CD Horizon Solera 4.75 mm Spinal System, used in AIS in our hospital, bearing Europe's regulations for introduction of medical implants in mind. The system consists of two cobalt chromium extra strong 4.75mm (CoCr Plus) rods, cobalt chrome multi-axial screws with dual lead thread form, standard laminar and pedicle hooks and modular crosslink plates.A correction rate of 51.5 % (48.0% - 54.9%) was found for the primary Cobb angle. These results are comparable with the range of correction rates (50% to 67.6%) as described in literature for similar instrumentation .A correction rate of 45.1% (39.3% - 50.9%) was found for the secondary Cobb angle. The results in literature vary between 43.7% and 63%. 3 studies show comparable results to ours and 4 studies show higher results compared to ours ,28,29.In some reports higher primary curve correction rates compared to this study, varying from 60 to 64% for the primary curve at two-year follow-up, were described, which might be explained by implant density, but these results are missing in these studies . In thisGenerally, a flattening of the thoracic spine has been reported in AIS patients We found nine complications in our study of which two (rod breakage) could potentially be attributed to the instrumentation. In total six revision surgeries were required in which total or part of the instrumentation was removed. Reasons for revision surgeries were rod breakage, invalidating pain, skin irritation, mental health problems, ventral screw protrusion and a deep wound infection. Two superficial wound infections and one rod breakage, which was coincidentally found, did not require revision surgery. Revision surgery rate in comparable literature ranged from 0% to 15.5% ,57,59,60The most common revision indications in posterior spinal correction and fusion in AIS are deep wound infection, pseudoarthrosis, pain, prominent implant and dislodged instrumentation In literature material breakage and dislodgement has been described to occur in 0.93% to 10.5% ,57,59,60This study was limited because of its retrospective design and its lack of a control group preventing direct comparison to other instrumentation devices. Study strengths are the relatively large sample size and the homogeneity of the group. Importantly, there was no conflict of interest by the operating surgeons or researchers. Future studies should contain a larger cohort, if feasible with a control group and longer follow-up of radiographs and prospective HRQoL questionnaires pre- and postoperatively. Preferably, clinical data should be gathered in a well-monitored and controlled setting before the introduction of a spinal device to the market.In patients with AIS the initial correction and maintenance of correction as achieved by posterior spinal fusion using the CD-Solera instrumentation is comparable to other reported devices. Complication rates are low and health related quality of life is comparable to literature. The CD-Solera can be regarded as a safe and effective instrumentation in surgical treatment of AIS.The authors declare that informed patient consent was taken from all the patients.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper


This paper reports a plasma treatment (PT) method for improving the surface hydrophilicity and mechanical properties of cellulose in reconstituted tobacco paper-base. The absorption and infiltration rates of water droplets on PT-reconstituted tobacco paper-base-15 s were significantly accelerated. Notably, the increased content of methylene and alkyl groups enabled the tobacco paper-base to absorb more useful substrates in the tobacco extract after plasma treatment. In addition, the tensile mechanical performance of reconstituted tobacco was significantly improved after plasma treatment, which indicated that the content of organic matter absorbed by the tobacco paper-base sheet was increased. Moreover, tobacco extract infiltrated on the surface of PT-reconstituted tobacco paper-base reached 37.7° within 30 s, while it reached 79.9° on the reconstituted tobacco paper-base. Finally, the mechanism by which the surface hydrophilicity and mechanical properties of the cellulose in the tobacco paper-base were improved is discussed. Chlorella strains by combining undirected mutagenesis and directional screening [With developments in paper treatment methodology, plasma techniques have attracted increasing attention for surface modification ,4,5,6,7.creening . Therefocreening . As a reLow-temperature plasma includes nitrogen, argon, ammonia, and oxygen plasma treatments. Due to the advantages of little damage, good treatment effects, energy conservation and environmental protection, oxygen plasma treatment has attracted attention for improving cellulose ,13,14,15Over recent decades, research on the use of reconstituted tobacco in paper-making has received much attention ,23,24. IA simple and cost-effective plasma treatment (PT) method was applied to improve the surface hydrophilicity and mechanical properties of reconstituted tobacco (PT-reconstituted tobacco). The hydrophilicity of the cellulose in tobacco paper-base was significantly improved under oxygen plasma treatment, and the infiltration rate of water droplets on its surface was clearly accelerated. Scanning electron microscope (SEM) analysis indicated the tobacco sheets still retained a consistent microstructure after oxygen plasma treatment. Furthermore, Fourier transform infrared spectrum and thermogravimetric analysis indicated that the PT-reconstituted tobacco absorbed more substrates in the tobacco extract. Moreover, the tension performance of PT-reconstituted tobacco was also greatly enhanced. The reconstituted tobacco paper-base sheets and the tobacco extract (liquid) were supplied by Anhui China Tobacco Reconstituted Tobacco Technology Co., Ltd., Hefei, China. To meet industrial production conditions, the tobacco extract was diluted by 50% in deionized water and then sonicated for 30 min. After that, the reconstituted tobacco paper-base was immersed in the tobacco extract for 30 s, and then placed in a 70 °C oven for 2 min. The obtained product was defined as reconstituted tobacco . The tensile test was conducted using a dynamic mechanical analyzer . Contact angles and the images of droplets were captured by commercial camera  and high-speed video camera . The rheological property of the tobacco extract was measured by a commercial rheometer . The surface of the tobacco paper-base was treated by plasma cleaning machine . The ultrasonic treatment was performed using an automatic ultrasonic cleaning machine  under 120 W working power and 25 °C temperature. The dimensions of the tobacco paper-base sheet for water/tobacco extract droplet infiltration experiments were 37.3 mm × 31.3 mm × 0.2 mm. In the quasistatic tensile test, the dimensions of the tobacco sheet were 20 mm × 4 mm × 5 mm for the central tensile area (ISO 37: 2005).The microstructures of the reconstituted tobaccos were observed by scanning electron microscope  under 3 kV. The infrared spectra of reconstituted tobacco paper-base, PT-reconstituted tobacco paper-base, reconstituted tobacco, PT-reconstituted tobacco, and tobacco extract were obtained using a Nicolet 8700 Fourier transform infrared (FT-IR) spectrometer . The test wavenumber range of tobacco sheet was from 4000 to 500 cmPlasma treatment changed the surface characteristics of cellulose in the paper-base sheets, thereby improving the infiltration properties of the tobacco sheets. SEM images provided detailed microstructure information of the traditional reconstituted tobacco paper-base and PT-reconstituted tobacco paper-base, showing the morphologies of the cellulose in tobacco paper-base before and after plasma treatment . Figure −1 and 1600 cm−1 indicate the presence of O−H bonds. The peak of PT-reconstituted tobacco paper-base at 2900 cm−1 shows the stretching vibration of C−H bonds saturated with methylene  . The recThe mechanical performance of the reconstituted tobacco paper-base and reconstituted tobacco was also explored . The ten−1 to 1000 s−1. The viscosity of the extract decreased with increasing in shear rate. The viscosities varied from 627.5 Pa·s to 4.8 Pa·s and from 388,793.1 Pa·s to 5.6 Pa·s under 25 °C and 70 °C, respectively. −1. The viscosity of the tobacco extract reached its maximum at 70 °C. In addition, the viscosity of the tobacco extract at 70 °C and shear rate of 10 s−1 was 476.7 Pa·s  of the PT-reconstituted tobacco was lower than that of reconstituted tobacco due to the greater absorption of organic matter. In addition, the O 1s and C 1s peaks in XPS spectra of the reconstituted tobacco and PT-reconstituted tobacco could be clearly observed, and the intensity of the C 1s peaks was enhanced in the PT-reconstituted tobacco spectra. Moreover, the maximum tension fracture force of the PT-reconstituted tobacco was increased by the plasma treatment. Moreover, the contact angle of the tobacco extract on the surface of the PT-tobacco paper-base dropped rapidly within 30 s, and the infiltration rate was greatly improved. In sum, the plasma treatment method shows significant potential for improving the quality of tobacco paper-base, and elsewhere in the paper-base industry.


Patients with a severe COVID-19 infection often require admission at an intensive care unit (ICU) when they develop acute respiratory distress syndrome (ARDS). Hyperinflammation plays an important role in the development of ARDS in COVID-19. Procalcitonin (PCT) is a biomarker which may be a predictor of hyperinflammation. When patients with COVID-19 are in the emergency department (ED), elevated PCT levels could be associated with severe COVID-19 infections. The goal of this study is to investigate the association between PCT levels and severe COVID-19 infections in the ED.This was a retrospective cohort study including patients with a confirmed COVID-19 infection who visited the ED of Erasmus Medical Center in Rotterdam, the Netherlands, between March and December 2020. The primary outcome was a severe COVID-19 infection, which was defined as patients who required ICU admission, all cause in-hospital mortality and mortality within 30 days after hospital discharge. PCT levels were measured during the ED visit. We used logistic regression to calculate the odds ratio (OR) with 95% confidence interval (95% CI) and corresponding area under the curve (AUC) of PCT on a severe COVID-19 infection, adjusting for bacterial coinfections, age, sex, comorbidities, C-reactive protein (CRP) and D-dimer.A total of 332 patients were included in the final analysis of this study, of which 105 patients reached the composite outcome of a severe COVID-19 infection. PCT showed an unadjusted OR of 4.19 (95%CI: 2.52–7.69) on a severe COVID-19 infection with an AUC of 0.82 (95% CI: 0.76–0.87). Corrected for bacterial coinfection, the OR of PCT was 4.05 (95% CI: 2.45–7.41). Adjusted for sex, bacterial coinfection, age any comorbidity, CRP and D-dimer, elevated PCT levels were still significantly associated with a severe COVID-19 infection with an adjusted OR of 2.11 (95% CI: 1.36–3.61). The AUC of this multivariable model was 0.85 (95%CI: 0.81–0.90).High PCT levels are associated with high rates of severe COVID-19 infections in patients with a COVID-19 infection in the ED. The routine measurement of PCT in patients with a COVID-19 infection in the ED may assist physicians in the clinical decision making process regarding ICU disposition.The online version contains supplementary material available at 10.1186/s12879-022-07144-5. Coronavirus disease COVID-19) caused by the novel Coronavirus SARS-CoV-2) was declared a pandemic on the 11th of March 2020 by the World Health Organization [9 caused  was declWhen patients with a COVID-19 infection present at the emergency department (ED), it is important to be able to identify patients who are at high risk of developing a severe COVID-19 infection. These patients may benefit from more extensive monitoring or early ICU admission.Different biomarkers have been identified as predictors of disease severity in COVID-19 , includiThe goal of this study is to investigate the association between PCT levels and severe COVID-19 infections in the ED.In this retrospective single center cohort study we included patients with a confirmed COVID-19 infection who visited the ED of Erasmus University Medical Center, in Rotterdam, the Netherlands, between 1 March 2020 and 31 December 2020. Erasmus University Medical Center is an academic hospital with 40.000 ED visits annually. This study was conducted in accordance with the Declaration of Helsinki . The institutional review board waivered informed consent for the retrospective use of clinical data of COVID-19 patients. The study was performed following the STROBE guidelines , if the COVID-19 infection was a secondary finding and not the primary reason for ED visit or when no follow-up data were available due to transfer to another hospital.Patient data including demographics, comorbidities, vital signs and laboratory tests were collected from the electronic patient records. Vital signs including heart rate, arterial oxygen saturation, respiratory rate, temperature and blood pressure were recorded from the time of ED visit. Laboratory testing during the ED visit included hemoglobin, red cell distribution width (RDW), leucocyte count, thrombocytes, total bilirubin, alanine aminotransferase (ALAT), aspartate aminotransferase (ALAT), lactate, D-dimer, C-reactive protein (CRP) and PCT. Patients were followed up for 30 days after hospital discharge. Patient disposition from the ED was categorized into discharge home from the ED, admission to general ward and admission to the ICU. Mortality data was classified as all cause in-hospital mortality and mortality at home within 30 days after hospital discharge. To correct for potential bacterial coinfections at ED visit, culture within 48 h of ED visit were reviewed. If blood, urine or sputum cultures or pneumococcal or legionella antigen tests were positive, patients were classified as having a bacterial coinfection.The primary outcome was a severe COVID-19 infection, defined as patients that were admitted to the ICU or patients that died by any in-hospital or within 30 days after hospital discharge. We calculated odds ratios (ORs) for PCT as continuous variable and commonly used cut-off values of PCT of 0.25 ng/mL, 0.5 ng/mL and 1.0 ng/mL. These cut-off values are derived from recommendations in previous literature .The secondary outcomes of this study were hospital admission, ICU admission and all-cause mortality, either in-hospital or within 30 days after hospital discharge. For the secondary outcomes, PCT was analyzed as both continuous variable and at the different cut-off values as previously described.PCT analysis was available as a standard laboratory test in patients who visited the ED with a suspected COVID-19 infection. Blood was collected in a lithium heparin tube and analyzed directly upon arrival in the clinical chemistry laboratory. PCT was measured using E801 Elecsys BRAHMS PCT reagent on a COBAS 8000 . The PCT values were available to the treating physician during the ED visit.Normally distributed variables were reported as mean with standard deviation (SD), non-normally distributed variables as median with interquartile range (IQR). Multiple imputation was used for handling missing data.Differences in dichotomous variables between the severe and non-severe COVID-19 infection patients were analyzed with a chi-square test. Differences in continuous variables were analyzed using an independent sample T-test for normally distributed data and a Mann–Whitney-U test for non-normally distributed data. Following, we did an univariate logistic regression analysis calculating the ORs of the variables that significantly differed between the groups.For the primary outcome, we calculated unadjusted odds ratios of PCT using univariate logistic regression analysis. We calculated adjusted ORs of PCT correcting for only bacterial coinfections and in a multivariable logistic regression model correcting for the following predefined variables: age, sex, bacterial coinfection, the presence of any comorbidity, CRP and D-dimer. We calculated receiver operating characteristic (ROC) curves and the corresponding area under the curve (AUC). The calibration statistics of the univariate analysis of PCT and the multivariable regression model were analyzed using the Hosmer-Lemshow test to investigate the goodness-of-fit and graphically reported by calibration plot with intercept and slope statistics.For the secondary outcomes, we used predefined cut-off values of 0.25 ng/mL, 0.5 ng/mL, 1.0 ng/mL of PCT and calculated the unadjusted ORs, the sensitivity, specificity, negative predictive value and positive predictive value on the primary and secondary outcomes of the study.The correct for the skewed distribution of PCT, we transformed PCT using the natural logarithm and performed the univariate analysis and calibration analysis in Additional file Statistical analyses were performed using ‘R’ version 4.1.1. We used the MICE package for multiple imputation of missing data and the RMS package for calibration statistics.Between 1 March 2020 and 31 December 2020, a total of 23,195 patients visited the ED of which 2188 were clinically suspected of having a COVID-19 infection. Of these patients, 477 patients had a positive COVID-19 PCR-test at the day of the ED visit. Procalcitonin was measured in 339 of these patients. Four patients were excluded due to an invalid procalcitonin measurement, two patients were excluded because the ED visit was not related to the COVID-19 infection and one patient was excluded because there was no follow up data available. A total of 332 patients were included in the analysis of this study  patients reached the composite outcome of a severe COVID-19 infection. Of these 105 patients, 44 (13%) were admitted to the ICU and 61 (18%) patients died. The non-severe COVID-19 infection group consisted of 227 (68%) patients, of which 67 (20%) were discharged directly from the ED and 158 (48%) were discharged from the general ward after an uncomplicated hospital admission. Baseline characteristics and differences between both groups including unadjusted ORs are shown in Table PCT showed an unadjusted OR of 4.19 95%CI: 2.52–7.69) on a severe COVID-19 infection with an AUC of 0.82 (CI: 0.76–0.87). Corrected for bacterial coinfection, high PCT remained associated with an increase in severe COVID-19 infections with an adjusted OR of 4.05 (95% CI: 2.45–7.41). In the multivariable model adjusted for sex, bacterial coinfection, age, any comorbidity, CRP and d-dimer, PCT was still significantly associated with a severe COVID-19 infection with an adjusted OR of 2.11 (95% CI: 1.36–3.61)  and the calibration plot showed a slope of 1.00 (95% CI: 0.64–1.42) and intercept of 0.00 (95% CI: − 0.42–0.48) , 12.12 (95% CI: 6.61–23.2) and 14.40 (95% CI: 6.45–36.77). The ORs of PCT at the different cut-off points of 0.25 ng/mL, 0.5 ng/mL and 1.0 ng/mL for the secondary outcomes are shown in Table In this study we found that elevated PCT levels are associated with an increase in the combined outcome of ICU admission and mortality. When treating a COVID-19 patient in the ED with an elevated PCT, the treating physician should not only look for possible bacterial coinfections but also be cautious of disease progression. Based on these findings, physicians should beware that patients with elevated PCT levels in the ED may have an increased risk of becoming severely ill. Although these findings need to be validated before PCT levels can be used to guide clinical decision-making on disposition of COVID-19 patients, PCT levels may serve as an early warning sign of possible clinical deterioration. Due to a low sensitivity of PCT on all outcomes, normal PCT levels should not be used in disposition decisions in COVID-19 patients in the ED.Our findings are supported by other studies, that showed that hospitalized COVID-19 patients with severe infections had higher PCT levels –17. OnlyPCT is primarily used as biomarker for bacterial infections. The findings of this study show a new use for PCT in the specific group of COVID-19 patients as marker of disease severity. Even after correcting for bacterial coinfections and inflammatory markers CRP and d-dimer, elevated PCT levels remained associated with a severe COVID-19 infection. Our finding that PCT is highly elevated in severe COVID-19 infections can be explained when looking at the pathways of synthesis of PCT . PCT synAt different cut-off values, PCT levels showed a high specificity for the hospital admission, ICU admission and mortality. On hospital admission, the lowest cut-off value of 0.25 ng/mL resulted in a specificity of 95%. This finding could be used as an argument to use PCT as support tool for hospital disposition decisions, because the majority of patients with a PCT above 0.25 ng/mL were admitted to the hospital. The sensitivity of PCT on all outcomes was low. Therefore, PCT is less suitable for being incorporated it in a rule out decision support tool of severe COVID-19 infections in the ED. Different cut-off values of PCT are recommended in distinguishing bacterial from viral infections, also ranging from 0.25 to 1.0 ng/mL . The OR A limitation of this study is its retrospective design. Patients were included if PCT was measured in the ED. PCT was only measured in patients where the primary reason of the ED visit was suspected COVID-19 infection. Patients that were initially not suspected of having COVID-19, but were diagnosed with a COVID-19 infection nonetheless, were not included in this study.In the secondary outcomes, the subgroups have a relatively small size, which influences the generalizability of the findings.The patient population consisted solely of confirmed COVID-19 patients. Therefore, the results are not biased by patients who visited the ED with a suspected COVID-19 infection but were diagnosed with an alternative diagnosis. However, the results of this study cannot be used for distinguishing a COVID-19 infection from other viral or bacterial infections .This study was performed in a cohort of patients who visited the ED in 2020, when no vaccinations against SARS-CoV-2 were available. It is unclear if these results are generalizable to the current ED setting, where the majority of patients is vaccinated .These reUsing only PCT as single predictor of the primary outcome showed a poor goodness-of-fit and the results may not be valid in other cohorts. Therefore, these results have to be validated in other cohorts. Because PCT has a skewed distribution of < 0.50 ng/mL in non-severe patients and from 0.50 up to 58 ng/mL in severely infected patients, the results may be biased by the patients with high PCT values. This can be corrected by using the natural logarithm of PCT, thereby reducing the range of PCT. The results were similar, but the goodness-of-fit was high (p = 0.91) .Additional file 2. Amount of missing data.Additional file 3. Univariate analysis of natural logarithm of procalcitonin on a severe COVID-19 infection.Additional file 4. Multivariable logistic regression model with different cut-off values of procalcitonin.


This study aimed to estimate the seroprevalence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection within the staff and student populations of the University of Corsica (France) during the second wave of the epidemic. Methods: A cross-sectional survey was conducted from 23 November 2020 to 31 January 2021. The participants underwent blood sampling using a fingerstick procedure and completed an anonymized questionnaire. Sera were tested for the presence of anti-SARS-CoV-2 IgG (ELISA-S) and, if positive, with an in-house virus neutralization test (VNT). Results: A total of 418 persons were included in the study. The overall seroprevalence was 12.8% , 9.8–16.6%). A total of 15 (31%) of the 49 individuals who had a positive ELISA-S also had a positive VNT. Seropositivity was associated with living at the city campus during the week and on weekends (OR = 3.74 [1.40–12.00]), using public transportation/carpooling (OR = 2.00 [1.01–4.02]), and being in contact with a person who tested positive for SARS-CoV-2 (OR = 2.32 [1.20–4.40]). The main symptoms associated with seropositivity were “having had an acute respiratory infection” (OR = 3.05 [1.43–6.43]) and “experiencing loss of smell” (OR = 16.4 [5.87–50.7]). Conclusion: These results could be useful for SARS-CoV-2 prevention and control on university campuses. Managing schools and universities during the pandemic of the coronavirus disease 2019 (COVID-19) has been a global challenge .In France, universities were closed during the first (March through May 2020) and second (30 October 2020 to 15 December 2020) COVID-19 waves. The events of 2020 and 2021 have shown that university campuses could pose significant challenges to the control of the spread of SARS-CoV-2 . A betteTo our knowledge, few studies have reported the seroprevalence of SARS-CoV-2 infection in universities ,6,7,8,9.In France, the second wave of COVID-19 occurred from the week of 24 August (2020w35) to the week of 21 December (2020w52), 2020. During this second wave, schools and universities were kept open because of expanded testing resources and the implementation of health protocols . Schools and universities opened at the beginning of September 2020 and remained open until 18 December 2020, with the exception of a 2-week autumn break. During this second wave, students at the University of Corsica had the opportunity to attend face-to-face classes once or twice per week. All staff were present on the university campus, with teleworking facilities (at least once a week).This was a university-population-based cross-sectional study that included voluntary participation by the students and staff of the University of Corsica, Corte, France. Corte, the only city campus of Corsica, has 7000 inhabitants. The staff group included administrative staff, teachers, researchers, and PhD students. The purpose of this study was to estimate the seroprevalence of IgG antibodies against SARS-CoV-2 from capillary blood samples collected from 23 November 2020 to 31 January 2021. In relation to the sampling, the participants were asked to complete a questionnaire investigating sociodemographic data, comorbidities, symptoms, and past history of COVID-19, as well as behavioral factors. Both the blood samples and the questionnaires were anonymized.n = 860) and students (n = 4442) of the University of Corsica were offered the possibility to participate in this study. They were invited by email to enroll in the study via their university mail on 23 November 2020. The students and staff were eligible to participate regardless of whether they had prior confirmed COVID-19 or COVID-19-related symptoms.All staff  self-reported SARS-CoV-2 testing history, as assessed using the following question: “Since January 2020, have you ever tested positive for a SARS-CoV-2 infection?” ; and (2) self-reported SARS-CoV-2 symptoms, as assessed using the following question: “Since January 2020, have you ever shown one of the following health statuses or symptoms?” ”, “influenza-like illness ”, “loss of taste”, “loss of smell”, “nausea and/or vomiting, diarrhea”, “abdominal pain”, or “other symptoms (that participants could specify)”. More than one answer was possible).Samples of capillary blood were obtained using a safety lancet on a cleansed finger puncture and collected into 0.8-mL tubes containing a coagulation activator and a serum separator. The tubes were centrifuged at 6000 rpm for 15 min and the resulting serum was stored at −20 °C until it was processed for serology.Samples were tested in duplicate for the presence of anti-SARS-CoV-2 IgG using the EUROIMMUN enzyme immunoassay kit for the semiquantitative detection of IgG antibodies against the S1 domain of the viral spike protein (ELISA-S) . The assay has a specificity and a sensitivity of around 99.8% and 90.3%, respectively, according to the manufacturer’s data.50) of the SARS-CoV-2 strain BavPat1 , and serial dilutions of serum (1/20–1/160) were used. Dilutions associated with a cytopathic effect (CPE) were considered negative , whereas those with no CPE at day 4 post infection were considered positive . The neutralization titer refers to the highest dilution of serum that yields a positive result. Specimens with a VNT titer of 40 were considered positive  kit . Quantification of S1-specific IgG was performed using a 6-point calibration curve covering a range from 1 to 120 relative units (RU)/mL. By multiplication with a factor of 3.2, results in RU/mL were converted into standardized binding antibody units (BAU)/mL. A result of <25.6 BAU/mL was considered negative and a result of ≥35.2 BAU/mL was considered positive. In all ELISA-S positive and borderline samples, neutralizing antibodies were also detected using a virus neutralization test (VNT), as described previously . VeroE6 positive .According to the overall IgG antibodies against SARS-CoV-2 seroprevalence observed in residual sera in mid-June in Corsica and at the end of October  , a minimp < 0.05.The primary outcome was the estimation of the proportion of participants with a positive ELISA-S test. Baseline characteristics and exposures were presented as the number (%) for factors and the mean or median for numerical variables, as appropriate. Univariate analyses were performed to measure the effect of each variable on the serological status. The Wilcoxon rank sum test was used to compare two independent samples. Chi-squared and Fisher’s tests were used to compare differences between groups for categorical variables. Statistical significance was set at Seroprevalence was initially calculated as the proportion of participants who had a positive ELISA-S result. Subsequently, considering the limitations of the ELISA-S method, the seroprevalence estimate was also adjusted based on test sensitivity (90.3%) and specificity (99.8%)  using Epitools software  .p ≥ 0.2) were retained for the multivariate analysis. Model selection was performed using the step Akaike information criterion. Two full models were built: one including the risk factors of exposure (such as type of accommodation), and the other including factors representing the consequence of the exposure . For the first type of model, seven variables  were selected to compose the full model. For the second type of model, four variables  were selected to compose the full model. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to explore factors potentially associated with a positive ELISA-S result.Univariate logistic regression models were built to identify the potential factors associated with the serological status. Only variables that were close to significance  .The study protocol was approved on 12 November 2020, by the ad hoc ethics committee (Comité de Protection des Personnes #2020-A00711-38). All data and questionnaires were checked and validated by the data protection officer of the University of Corsica. All participants in this study were volunteers and unpaid. The entire university population was informed that the samples would be used for epidemiological studies via an information letter. Written informed consent was required for the use of the data in the analyses.n = 271) were female and the median age was 31 (IQR: 22–44) years. Half of the participants  were postgraduate. Of the 418 participants, 44% (n = 184) lived in Corte (the city hosting the university campus) and about half of the population lived in apartments . Finally, 28% (n = 117) of the participants had at least one comorbidity  who filled out a questionnaire and had full serology were included in the study . The basorbidity .n = 149) of the participants declared having had COVID-19-related symptoms since 1 January 2020, and 6.5% (n = 27) of them declared having previously tested positive (PCR test) for SARS-CoV-2 since 1 January 2020. Among the 27 confirmed cases, 5.3% (n = 22) declared having had at least one symptom related to virologically confirmed SARS-CoV-2.As shown in n = 109 out of 418) of the participants reported having had at least one contact with a confirmed COVID-19-positive individual since 1 January 2020. Among those 109 participants, 28% (n = 31) had more than one contact episode. The majority of contacts consisted of interactions with coworkers or classmates , followed by contact with family members  and friends .Twenty-six percent (p = 0.57)  tested positive on ELISA-S, with similar values recorded between students (10.5%) and staff members (12.4%) ( = 0.57) . Accordi = 0.57) .The serological status and the self-reported positive SARS-CoV-2 test results according to the presence or absence of symptoms are presented in n = 31) declared having experienced at least one COVID-19 symptom since 1 January 2020. The symptoms experienced by participants that were most associated with a positive ELISA-S result (p < 0.001) were loss of smell  and loss of taste .Among the 49 participants with a positive ELISA-S result, 63.3% (n = 38 out of 49) of the ELISA-S positive samples were measured quantitatively using QuantiVac ELISA-S. The mean IgG antibody level of symptomatic individuals (124.37 BAU/mL) was slightly higher than, but not significantly different from, that of asymptomatic individuals, at 62.33 BAU/mL (p = 0.08175) (The IgG antibody levels against SARS-CoV-2 in 77.5% (0.08175) .n = 15) had a positive result on the VNT. Among the 15 participants who were seroneutralization positive, 46.7% (n = 7) had a VNT titer of 40, 33.3% (n = 5) had a VNT titer of 80, and 20.0% (n = 3) had a VNT titer of 160.The seroneutralization results obtained for the study population are presented in p = 0.024 and p < 0.001, respectively)  .n = 249) of the participants reported never using public transportation or carpooling. Regarding social interactions outside of a work, home, and public transport context , 44% (n = 184) of the participants reported having a few of these interactions several times per week. Regarding the adherence to barrier behaviors, 68% (n = 284) of the participants mentioned adopting the correct actions and behaviors to protect themselves and others from SARS-CoV-2. At least 30% (n = 152) of the respondents reported being a little concerned about the consequences of SARS-CoV-2 infection on their own health, whereas 43% (n = 178) were very concerned about the health of individuals close to them.The sociobehavioral characteristics of the study population are presented in p = 0.014), using public transportation/carpooling , having a case contact since January 2020 , the presence of an acute respiratory infection  and loss of smell . Living in a house  was a protective factor against prior SARS-CoV-2 infection , showed that 12.8% of the participants who had a positive ELISA-S result among the students and staff had evidence of prior infection, with at least 30% of them showing neutralizing antibodies. Notably, this rate, which was similar between the students and faculty staff, was significantly higher than the 8.04%  observed in the residual sera of the Corsican general population during the same period, as assessed using the same serological assays . In thisHowever, our seropositivity rate was in the lower range of that observed in a cross-sectional UK study of SARS-CoV-2 antibody seroprevalence among the university student population, which reported a rate ranging between 7.6% and 29.7% across the five participating UK universities during the second wave of the epidemic (December 2020) .We investigated the association between demographical, clinical, and lifestyle factors and seropositivity. In the present study, participants living in Corte (city campus) during the week and on weekends were at greater risk than were participants living outside Corte during the week and outside Corte on weekends. Because Corte is a small town (approximately 7000 inhabitants), we can assume that contacts between people are more frequent, as they come into contact more often in the same indoor places . Furthermore, we know that SARS-CoV-2 can remain airborne for up to three hours and survive on surfaces . In agreIn addition, we investigated the association between seropositivity and a history of SARS-CoV-2. Among the participants, a positive ELISA-S result was significantly associated with case contact or with having reported testing positive for SARS-CoV-2 since January 2020. The majority of case contacts reported by the participants consisted of interactions with coworkers or classmates, which suggests that the university community may favor the transmission of and contamination with SARS-CoV-2. This is in line with previous studies showing that close contact with people infected with COVID-19 increases viral transmission .Furthermore, seroprevalence was strongly associated with at least one of the symptoms self-reported since 1 January 2020. The main symptoms associated with seropositivity were “having had an acute respiratory infection” and “experiencing loss of smell”. As reported previously , anosmiaInterestingly, almost 40% of the participants who were positive on ELISA-S reported no symptoms since January 2020. This is in agreement with current data suggesting that infected persons without symptoms—including both presymptomatic and asymptomatic persons—account for more than 40% of all SARS-CoV-2 transmission ,28,29. TThis study had several limitations. The selected university community may not be representative; indeed, our population is composed mainly of university staff  since this is the group that responded most favorably to the call for participation. It is a specific population and data were obtained through a self-reported questionnaire; however, it is reasonable to assume that the awareness and memory of symptoms that may be related to COVID-19 have increased, leading to a lower estimate of the asymptomatic fraction. Furthermore, these results cannot be extrapolated directly to the general population. In turn, this study had several strengths. In particular, it was conducted among a university community for which data were scarcely available. In addition, serological samples were collected during the second wave of intense SARS-CoV-2 circulation. Finally, several serological methods were used to improve the interpretation of the seroprevalence results.This study revealed that the university community showed a significantly higher seroprevalence relative to the general population and that a large proportion had no significant symptoms related to COVID-19. The results highlight the setting in which adherence to measures including hand and respiratory hygiene, physical distancing, mask wearing, and the ventilation of indoor environments is critical to control the spread of SARS-CoV-2 viruses. It would be interesting to carry out a follow-up antibody quantification to evaluate the natural immunity and/or vaccination of the university population over time.


Bleeding is a common complication of extracorporeal membrane oxygenation (ECMO), leading to increased mortality. Since one of its main complications is bleeding, platelet transfusions are frequently prescribed for children on ECMO. However, there is currently very little information on the effect of platelet transfusions on the function of the ECMO oxygenator. Our objective was to describe the effect of platelet transfusions on oxygenator function.<18 years) who received ECMO support in our pediatric intensive care unit (PICU) between January 2017 and December 2019. Oxygenator function, measured before and after platelet transfusion, was assessed by post-oxygenator PECMOO2 and the gradient in pre- post-oxygenator pressures (Delta Pressure).In this retrospective study, we included all children (p = 0.69) and post-membrane PECMOO2 was 6 mmHg (p = 0.49). However, in the subgroup with the lowest quartile of pre-transfusion oxygenator function, the post-transfusion change in delta-pressure was −5.2 ± 2.7 mmHg (p < 0.001) and the post-transfusion change in PECMOO2 was −118 ± 49 (p < 0.001). The area under the ROC curve for the pre-transfusion delta-pressure and PECMOO2 to predict a worsening of the oxygenator function were 0.72 (95%CI 0.63–0.81) and 0.71 (95%CI 0.64–0.78), respectively. Using regression models, pre-transfusion delta-pressure and PECMOO2 were the only independent factors associated with oxygenator function worsening (p < 0.001).Over 3 years, we analyzed 235 platelet transfusions from 55 children who received ECMO support. Thirty-two (80%) of children were on veno-arterial ECMO and majority of them were peripherally cannulated. When looking at all transfusions, the post-transfusion change in delta-pressure was 0.1 mmHg (Our study suggests that overall, platelet transfusions do not seem to impact the ECMO oxygenator’s function. However, in the subgroup of patients with the lowest pre-transfusion oxygenator function, platelet transfusions were independently associated with a worsening function. Future studies should investigate if this warrants adjustments of the anticoagulation strategy around the platelet transfusion, especially among patients with lower oxygenator function. Extracorporeal membrane oxygenation (ECMO) is a temporary support measure used in children with refractory cardiorespiratory failure. However, bleeding and thrombosis remained frequent complications. Dalton et al. reported bleeding events in 70% of children on ECMO and thrombotic events in 38% . These aOn ECMO, blood exposure to the artificial surface of the circuit, inflammatory response, and shear stress-induced platelet activation are the main reasons for a consumptive coagulopathy, which is believed to play an important role in the increased risk of thrombosis and bleeding , 3. QualSome underlying mechanisms that could explain worse clinical outcomes after platelet transfusions have been explored. The activated platelets during the ECMO support seem to result in excessive circuit clot burden, especially in the oxygenator membrane , which iThis retrospective study was conducted at a quaternary care university hospital. Following our institutional review board approval , the data was extracted from electronic medical records. All children 0–18 years of age admitted to our pediatric intensive care unit (PICU) between January 2017 and December 2019 were eligible. We included all children who received ECMO support, irrespective of the indication. We did not include patients undergoing ECMO in other units, such as the neonatal intensive care unit or the adult intensive cares. We identified ECMO procedure codes in the EMR using the International Classification of Diseases Tenth Revision Clinical Modification (ICD10-CM) code 5A15223.9/L or irrespective of platelet count if the patient is with clinical bleeding.Extracorporeal membrane oxygenation system used during the study period remained consistent with Quadrox-D oxygenator, Rotaflow centrifugal pump , and no bladder or reservoir. An oxygen air blender was used for gas exchange across the oxygen membrane. The oxygenator heat exchanger was set at 37°C. The anticoagulation practices also remained consistent during the study period. The institutional anticoagulation protocol was followed with unfractionated heparin as a primary anticoagulant. Coagulation status was monitored every 4–8 h. The goal heparin assay (anti-Xa) was 0.3–0.7 U/mL and the aPTT goal was 1.5–2 times the control value in seconds. If there was a discrepancy between the aPTT and anti-Xa, we follow the anti-Xa level. In addition, Thromboelastogram (TEG) was performed at least daily. Per our institution ECMO policy, we transfuse platelets if the platelet count is less than 100 × 10We collected demographic, clinical, laboratory, platelet transfusion, and ECMO circuit variables. We measured the bleeding and thrombotic event as reported in the EMR. Bleeding was defined as per extracorporeal membrane oxygenation support (ELSO) guidelines based on the red blood cell transfusion or other intervention such as surgical or endoscopic intervention . ThromboECMOO2).The primary outcome was the change in oxygenator function, as evaluated by the change in oxygenator membrane pressure gradient (delta-pressure), using the difference between the pre- and post-oxygenator pressures. The secondary outcome was the pre- post-transfusion change in post-oxygenator oxygen partial pressure .To identify factors associated with worse post-oxygenation oxygenator function, we categorized the platelet transfusion events according to the 75th percentile of the change in delta-pressure (≤2 mmHg vs. >2 mmHg) and change in Pn) and percentages; continuous data were reported as median and interquartile ranges (IQR). We used the paired Student T-test to evaluate the significance of PECMOO2 and Delta-Pressure changes, before and after platelet transfusion, since the distribution was normal. We used Wilcoxon Rank Test to evaluate differences among groups, as the distributions were not normal. Multivariable linear regression models were developed to evaluate the post-transfusion change in PECMOO2 and Delta-Pressure, as well as the cumulative effect of transfusions. Variables were considered potential predictors if they were associated with the outcome in univariate analysis (p < 0.10). The final model was selected using a bidirectional step-wise selection on the potential predictors with a significance criterion of the p-value of <0.05 to enter in the model and a p-value of >0.1 to be removed. No variables other than the PECMOO2 and Delta-Pressure were forced into the models. We evaluated the accuracy to predict a worsening of the oxygenator function with an area under the receiver operating characteristic (AUC). AUC was considered excellent if >0.90; good between 0.80 and 0.90; fair between 0.70 and 0.79; poor between 0.60 and 0.69; and not discriminatory <0.60 .The categorical variables were reported as counts  received at least one platelet transfusion, for a total of 235 transfusions. Of these, 22 (55%) were males, with the median age of 4 months (IQR 0.45–33). As shown in 9/L (IQR 69-155). The median transfused platelet volume was 10 mL/kg (IQR 10-10).As presented in p = 0.69, p < 0.001, The mean oxygenator delta-pressures were 14.6 ± 9.3 mmHg (before platelet transfusion) and 14.8 ± 9.5 mmHg (after platelet transfusion). The median change in delta-pressure was 0 mmHg (IQR −1 to 2) mmHg, which was not a statistically significant difference from pre-transfusion baseline (p < 0.04).The area under the ROC curve for the pre-transfusion delta-pressure to predict a worsening of the oxygenator function was 0.72  and 366 ± 132 mmHg (after platelet transfusion). The median change in post-membrane PECMOO2 was 6 mmHg (IQR −45 to 50), which was not a statistically significant difference from pre-transfusion baseline  was the pre-transfusion PECMOO2  had significantly worse post-transfusion PECMOO2, when compared to patients with PECMOO2 above this cutoff .The area under the ROC curve for the pre-transfusion Pp = 0.57) or PECMOO2 (p = 0.68).When adjusting for the pre-transfusion oxygenator function, there was no cumulative effect of the number of transfusions on post-transfusion change in delta-pressure . It has been reported that immediately following ECMO support initiation, neonates drop their platelet count by 25–60% and children by 30%, due to hemodilution and platelet destruction by the pump . Written informed consent from the participants’ legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.All authors listed have made a substantial, direct, and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Purpose: This study aimed to explore a predictive risk-stratification model combing clinical characteristics and lipid profiles in multiple myeloma (MM) patients.Methods: The data of 275 patients in Sun Yat-Sen University Cancer Center were retrospectively analyzed and randomly divided into the training (n = 138) and validation (n=137) cohorts. Triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), lactate dehydrogenase (LDH), Apolipoprotein B (Apo B) and Apo B/Apolipoprotein A1 (Apo A1) ratio were the prognostic factors identified through univariate and multivariate Cox analysis.Results: A 6-prognostic factor model was constructed based on Lasso regression. Patients were divided into low- and high-risk groups and the former group showed longer overall survival (OS) time (p<0.05). The area under the curve (AUC) of the risk score model for 5-and 10-year OS were 0.756 [95% CI: 0.661-0.850] and 0.940 [95% CI: 0.883-0.997], which exhibited better accuracy than International Staging System (ISS) and Durie and Salmon (DS) stage.Conclusion: This study aims to combine the lipid metabolism profile with the clinical characteristics of MM patients to generate a prognostic model. The nomogram integrating ISS stage and risk score increased the prediction accuracy. This model can monitor lipid profile as a simple and effective method, which has certain clinical significance for improving the accuracy of the prognosis and exploring potential therapeutic targets. Multiple myeloma (MM) is a cytogenetically heterogeneous plasma cell clonal proliferative disease that accounts for around 17% of all hematological malignancies Several current prognostic evaluation systems have been widely used in the clinic. The Durie and Salmon (DS) system was first proposed in 1975 Recent studies have indicated that the levels of serum lipids play an important role in tumor development and metastasis and have prognostic value In this study, we aimed to clarify the relationships between lipid and apolipoprotein profiles and the types and stages of MM. We retrospectively analyzed the lipid and apolipoprotein profiles of patients with MM and explored their predictive values when combined with existing prognostic indicators and clinical pathology. We developed a model based on lipid profiles that has high prognostic accuracy. Our data support the inclusion of lipid to improve the predictive accuracy of clinically established biomarkers in the prognosis of MM.The study was a retrospectively one, approved by the Clinical Research Ethics Committee of Sun Yat-sen University Cancer Center. We analyzed the clinical data of 372 MM adult patients (age ≥18 years) who were pathologically diagnosed and previously untreated in the Sun Yat-sen University Cancer Center (SYSUCC) between January 2007 and September 2019. The exclusion criteria were described as following: a) patients diagnosed with undefined significance, such as smoldering multiple myeloma, primary amyloidosis or solitary extramedullary plasmacytoma, b) patients previously taken or regular intake of any lipid regulating agent, without normal liver and kidney functions, c) patients lacking of data concerning lipid profile data, d) patients without complete follow-up data or died within a month, e) patients with earlier or simultaneously diagnosed other malignant diseases. Finally, a total of 275 patients were eligible for analysis in this study. And the patients were then randomly divided into the training (n = 138) and validation (n = 137) cohorts.The classical drugs-based regimens given to patients included VAD , DVD  The data of each enrolled patient was acquired after diagnosis and before the initiation of treatment. Baseline clinical characteristics involved age, gender, height, weight, albumin (ALB), serum creatinine (SCr), calcium (Ca), lactate dehydrogenase (LDH), β-2 micro globin (β-2 MG), DS stage, ISS stage, date of diagnosis, date of death or last follow-up, along with initial therapy and response of treatment. While the lipid profiles contained apolipoprotein B (Apo B), apolipoprotein A1 (Apo A1), cholesterol (CHO), triglycerides (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL). By the weight (in kilograms) divided by height (in meters) squared calculated body mass index (BMI). The laboratory tests were conducted on fresh blood samples collected after overnight fasting in the laboratory of the Sun Yat-san University Cancer Center. The median time interval between blood sampling and the commencement of treatment was 14 days (3-25 days) and the time interval was not associated with survival.Patient follow-up was performed by hospital outpatient follow-up or telephone interviews every 6 months for the first 3 years and then every year to determine recurrence or death. The last follow-up date was November 31st, 2020, to confirm the final conditions of recruited patients. The main outcome of the study was overall survival (OS) which was defined as the time interval between the date of diagnosis and date of death for deceased patients or the date of the last follow-up.20 was used to determine the optimal cutoff values based on OS for each of potential prognostic biomarkers among lipids and apolipoproteins profiles and stratify patients into low- and high-level sub-groups. Univariate and multivariate Cox analyses were used to evaluate the individual prognostic values of the profiles. The optimum cut-off values for all variables were determined using the X-tile software as follows: Age (60 year), BMI (22), ALB (30 g/L), SCr (81 umol/L), Ca (2.5 mmol/L), ApoB (1.1 g/L), ApoA1 (1.05 g/L), ApoB/ ApoA1 (1.09), TG (1.25 mmol/L), LDH (215 U/L), CHO (3.2 mmol/L), LDL (1.9 mmol/L), HDL (0.9 mmol/L) and β-2 MG (5 mg/L).The X-tile software (3.6.1)P-value threshold of < 0.05. R software  was used to perform the statistical analyses.A one-way ANOVA was used to analyze continuous variables and the Chisq test or Fisher exact test for categorical variables via the SPSS software version 25 . Continuous factors of the patients, such as demographic distributions, distribution characteristics of the lipids level and the clinical variables were expressed as the mean ± standard deviation. The X-tile software was used to determine the optimal cut-off values for dividing the continuous variables into categorical variables. Univariable and multivariable Cox regression hazard models were employed to identify independent prognostic factors of OS for MM according to their cut-off values. Next, the least absolute shrinkage and selection operator (LASSO) regression analysis was used to explore the best weighting coefficients of these independent prognostic factors. This approach is an established method for the regression of high-dimensional data. Survival analyses were performed using the Kaplan-Meier method. The prognostic accuracy of the model was quantified using time-dependent receiver operating characteristic curves (ROC) and the area under the curve (AUC). The analyses mentioned above were first performed in the training cohort, and then validated in the validation cohort. All statistical tests were two-sided and differences were significant at a Figure 1. The baseline patient characteristics are summarized in Supplemental Table 1. The age of patients ranged from 24 to 84 years with a median age of 60 years. 164 patients (59.6%) were male and 111 (40.4%) patients were female. Approximately half of the patients had a serum monoclonal protein of IgG, while about 22.9% (63) patients with monoclonal protein of IgA and 22.1% (61) patients did not have records of monoclonal protein. In total, 96 (34.9%) patients received bortezomib-based regimens and 179 (65.1%) patients received classical drugs-based regimens. All patients were given at least four cycles of treatment, 18 (6.5%) patients followed by ASCT. Approximately 25% of cases achieved complete response (CR).A total number of 275 MM patients were enrolled with available lipids profiles and survival data and randomly divided them into the training cohort (n=138) and validation cohort (n=137) as shown in Figure 2, P<0.05).The univariate and multivariate COX analyses were used to evaluate the individual prognostic values of the profiles. Univariate Cox analysis showed that lipids including ApoB, CHO, TG, LDH, and HDL were independent prognostic indicators (P<0.05). While multivariate analysis showed that variables including ApoB, TG, LDH, LDL, HDL and the ApoB/ApoA1 ratio were independent prognostic indicators . The cumulative OS rate of patients in the low HDL, CHO, TG, ApoB group was significantly lower than that of patients in the high group (P<0.05) . Besides, the OS rate of patients in the low LDH, β2MG group was significantly higher than that of patients in the high group (P<0.05) .In addition, we compared the OS between the low- and high- level of the lipids and apolipoproteins with Kaplan-Meier survival analysis respectively . The equation for the model was as follows:The potential prognostic factors based on univariate and multivariate Cox analysis were considered to associate with OS. Then, the LASSO regression analysis was employed to construct a prognostic model using the seven prognostic factors  in the training cohort. Based on the penalized maximum likelihood estimator of 1000 bootstrap replicates, a 6-prognostic factor model was established via the minimum criteria optimal λ value (Table 1).The lipid profile risk score was generated for each patient according to the above formula. Patients were further grouped into high and low-risk groups according to the median threshold of the lipids profile risk scores based on the training cohort. The distribution of age, ApoB, TG, HDL, LDH, β2MG and ISS stage were significantly different between the two subgroups  in the training cohort . The AUCs for 1-, 3-, 5- and 10-year survival were 0.696, 0.705, 0.756 and 0.940, respectively . Moreover, it could be seen that treatment response has correlation with risk levels, as shown in Supplemental Figure 2. The low-risk group had longer OS time in classical drugs-based regimens and bortezomib-based groups. And the AUCs for 1-, 3-, 5- and 10-year survival showed certain predictive significance.The survival of MM patients is summarized in Figure ). The AUCs for 1-, 3-, 5- and 10-year survival were 0.707, 0.734, 0.661 and 0.806 . We analyzed the distribution of the lipid profile risk scores in patients with different survival outcomes using dot plots to compare the survival of subjects. Our data showed that the survival of patients in the low-risk group was higher than survival in the high-risk group  in the two cohorts.Furthermore, in validation cohort, the low-risk group had longer OS time (p<0.05) as well  in the training and validation cohorts. After adjusting other clinical confounding factors in multivariate Cox analysis, the risk score was still an independent prognostic factor for OS  in the two cohorts mentioned above.The univariate and multivariate Cox analyses for risk score and other prognostic values were performed in MM. The univariate analysis indicated that the risk score was an independent prognostic indicator for OS . The AUC of the risk score model for 3-year OS was 0.661 [95% CI: 0.544-0.778]in the validation cohort . The AUC of the risk score model for 10-year OS was 0.940[95% CI: 0.883-0.997] , while the β2-MG, ISS and DS stage models were 0.545 [95%CI: 0.338-0.756], 0.519 [95%CI: 0.272-0.765] and 0.410 [95% CI: 0.273-0.582], which was significantly higher than the AUCs for ISS and DS stage models. The AUC of the risk score model for 10-year OS was 0.806 [95% CI: 0.689-0.923]  in the validation cohorts. Heat maps were used to compare the clinical characteristics and level of serum lipid profiles. Patients with high-risk scores were associated with older age and a higher stage of disease .The AUC of the risk score model for 5-year OS was 0.756 [95% CI: 0.662-0.850]. The AUCs for the level of β2-MG, ISS and DS stage models were 0.656 [95%CI: 0.562-0.750], 0.637 [95%CI: 0.536-0.738] and 0.641 [95% CI: 0.538-0.745] in the training cohort. However, there were no significant differences in prognostic accuracy was observed between the three models  combining two cohorts. The calibration plots showed good performance of the nomogram in predicting the 1, 3, 5, 10-year OS . The AUC of merged score for 1, 3, 5, 10-year were 0.725 [95% CI: 0.624-0.827], 0.709 [95% CI: 0.642-0.777], 0.732 [95% CI: 0.660-0.803] and 0.857 [95% CI: 0.767-0.948], which was significantly higher than ISS stage, suggesting that the nomogram can enhance the OS prediction compare to the standard prognostic factor .Furthermore, to generate a more accurate evaluation system, a nomogram was used to integrate the classic prognostic factors, the ISS stage  and can be used to stratify MM patients into high and low-risk groups that had significant differences in OS. Also, a nomogram was developed to predict survival and the model was tested with the time-dependent ROC curve. Our results indicated that the model had improved prognostic value compared to the other staging systems.The relationship between lipids and apolipoproteins in MM has not been systematically investigated and few comprehensive studies have reported on the clinical significance of lipid profiles in MM. To the best of our knowledge, no previous study has combined lipids and apolipoproteins as potential prognostic indicators in patients with MM. In this study, we analyzed the lipids and apolipoprotein levels of MM patients using a multi-layered approach. We found that most circulating lipids and apolipoproteins levels in MM patients are related to survival. We also retrospectively analyzed the relationship between risk level and clinicopathological features, treatment and prognosis. In the present study, 179 patients were treated with classical drugs-based regimens and 96 patients received bortezomib-based regimens. It was found that both in two subgroups, patients in low-risk group had longer OS time. Moreover, combined ROC analysis, our model showed it also has certain predictive significance for the survival of high- and low-risk patients in subgroups. In these cases, the lipid profiles are related to the etiology and prognosis of MM. Lipid profiles and several parameters identified as having prognostic value In MM, precision medicine approaches are required along with an improved understanding of the role of metabolism in tumorigenesis ApoB levels directly correlate with circulating serum cholesterol levels which can transport lipids to cells in the human body in vivo and in vitro studies have found that statins, which are HMG-CoA reductase inhibitors used for the treatment of hyperlipidemia, may increase the susceptibility of myeloma cells to apoptosis through a variety of ways. Reduce the mortality of MM to a certain extent, and may increase the sensitivity and resistance of treatment In this study, we aimed to assess lipid levels and the relationship between these levels and prognosis of patients with MM. Abnormal lipid metabolism is an important feature of tumor cells and is also a potential target for tumor treatment As mentioned above, the significance of lipid parameters in predicting survival has been confirmed. However, there is no relevant literature that can improve the prognosis of MM after adjusting lipid metabolism while most of the effects of lipid metabolism on tumor prognosis are based on retrospective data analysis to establish predictive models, and only theoretically study the effect of lipid metabolism on prognosis. Retrospective data are mostly cross-sectional studies, short-term follow-up and heterogeneity seemed to be a limitation. Moreover, simple dyslipidemia is difficult to diagnose and rarely treated, and the patients are different from the background population at baseline and have multiple confounding factors. The direction of the relationship between tumor development and changes in blood lipid levels cannot be determined by the results of such studies alone. We look forward to the follow-up study of the interaction between lipid metabolism and tumor prognosis from the molecular mechanism level, which may provide certain help for the whole-process management and prognosis prediction of patients from the perspective of lipid metabolism.Further research is needed to explore the role of lipids and apolipoproteins in MM. The expression of lipid profile levels in the serum exists for a short time but reflect the overall health status of the patient and liver metabolism function as a whole. This approach is clinically significant and can be used to detect the condition of patients and inform decision-making in the clinic Whilst our study provided intriguing research findings, it also has several limitations. Firstly, the data collection period was long and it is impossible to systematically compare it with a certain evaluation system as a whole. Secondly, all patient data were collected from a single cancer center which may introduce confounding factors. The sample size was limited and further research is needed to determine if the optimal critical values can be used in a larger range. It is expected that further large-scale, prospective, multicenter studies are needed to validate our findings. Finally, the potential mechanisms of lipid metabolism in the occurrence and development of MM remain to be fully determined.We generated a lipid profile-based model to predict the prognosis of MM patients. Lipid profiles are novel prognostic biomarkers that have high accuracy in predicting survival and may inform the future development of precision medicine in patients with MM.Supplementary figures and table.Click here for additional data file.


Pseudomonas aeruginosa infection model, aiming to draw attention to the appropriate selection of solvents for difficult-to-solubilize anti-infectives.Dimethyl sulfoxide (DMSO) and polyethylene glycols (PEGs) are frequently used as potent excipients in pharmaceutical formulations. However, these agents also have an interesting antimicrobial and anti-inflammatory profile that could interfere with the efficacy testing of anti-infective compounds when the latter are solubilized in DMSO or PEGs. Here, we demonstrate the antimicrobial and anti-inflammatory effects of DMSO-PEG400 in a murine IMPORTANCE Our study demonstrates the antimicrobial and anti-inflammatory effects of the combination of DMSO and PEG400 against Pseudomonas aeruginosain vitro and in vivo in a murine infection model of heightened intestinal permeability. The aim of this study is to draw attention to the appropriate selection of solvents for difficult-to-solubilize anti-infective compounds, to avoid interference with the assay or system tested. This is an extremely important consideration, since potential antimicrobial and anti-inflammatory effects of the solvent vehicle are detrimental to research studies on the efficacy of new anti-infective agents, given that the vehicle effect can mask the effect of the tested compounds. Our results can therefore be of great value to the scientific community, as they can guide researchers in the future to avoid this significant pitfall that can cost substantial amounts of money and valuable time during investigations of the effects of novel, difficult-to-solubilize antimicrobial compounds. They have frequently been used to improve the solubility of compounds that are difficult to solubilize in water or aqueous solvents (–4–https://www.dmso.org/articles/information/herschler.htm) and anti-inflammatory properties (Dimethyl sulfoxide (DMSO) and polyethylene glycols (PEGs) have long been used as versatile excipients in the formulation of pharmaceutical agents   that target bacterial virulence  (600) of 0.01 and were used as starter cultures, which were incubated . Their OD600 was measured at different time points, and the growth curve was plotted using GraphPad Prism . To understand the DMSO-PEG400 effect on PA growth, we interrogated how it affects bacterial growth oratory) ; CF1, a in vitro was already available in the literature (8% DMSO)  were added in different tubes with starter culture. The tubes were incubated for 24 h , and following incubation, the culture OD600 was determined. Based on this assay, the MIC for PEG400 was determined to be the 35% concentration.The MIC of DMSO for PA 8% DMSO) . We therin vivo studies, 10-week-old male C57BL/6 mice (The Jackson Laboratory) were maintained in a specific-pathogen-free environment at the Massachusetts General Hospital (MGH), in a 12-h-light–12-h-dark photoperiod, at an ambient temperature of 22 ± 1°C, with food and water access ad libitum. Animal protocols were approved by the Institutional Animal Care and Use Committee (IACUC) at MGH (protocol no. 2006N000093) and are in strict accordance with the guidelines of the Committee on Animals of MGH, Harvard Medical School, and the regulations of the Subcommittee on Research Animal Care of MGH and the National Institutes of Health.For the Mice were anesthetized , and the dorsal fur was removed with a clipper. A 30% total body surface area (TBSA) dorsal burn was induced by immersion in 90°C water for 8 s, using a polystyrene foam template, as described in reference 4 containing approximately 3 × 105 CFU of PA14 was intradermally injected at the burn eschar in the burn-plus-infection (BI) group. Mice in the no-infection group received 100 μl of 10 mM MgSO4. For the preparation of every 1,000 μl of the 5% DMSO–40% PEG400 combination, 550 μl of distilled water, 50 μl of DMSO  and 400 μl of PEG400  were mixed in a sterile tube. One hundred microliter of 5% DMSO–40% PEG400 was administered subcutaneously at 1, 3, 5, 7, 11, 15, and 19 h post-BI.  Every group consisted of 5 mice. Experiments were repeated twice.Bacteria were grown in LB broth or LB agar plates. One hundred microliters of 10 mM MgSOg; 90 s) and was used for FITC level assessment . The BI mice exerted a higher FITC-dextran flux out of the intestinal lumen  than the burn-alone group . DMSO-PEG400 significantly reduced the FITC-dextran flux , indicating mitigation of the bacterially mediated derangement of the intestinal permeability  of fluorescein isothiocyanate (FITC)-dextran  in phosphate-buffered saline (PBS). At 22 h post-BI, mice were euthanized. Blood samples were collected via aseptic cardiac puncture in BD Microtainer SST amber blood collection tubes. Serum was extracted by centrifugation . DMSO-PEG400 treatment conferred a significant TNF-α reduction . Likewise, DMSO-PEG400 administration also reduced the IL-6 levels in the ileum  (P < 0.05)  . Finally < 0.05) , were al < 0.05) .P < 0.001), indicating an antibacterial role for this solvent combination.Subsequently, we tested the effect of DMSO-PEG400 in the post-BI bacterial dissemination from the burn site to the spleen. Spleen samples were aseptically harvested through midline laparotomy and were homogenized in 1 ml sterile PBS using a tissue homogenizer . The homogenates were serially diluted 1/10 to 1/1,000 and plated on LB agar plates. Plates were incubated at 37°C, and CFU were quantified. post hoc comparisons was used for all statistical analyses (GraphPad Prism). P values of <0.05 were considered statistically significant.One-way analysis of variance (ANOVA) with Tukey’s in vitro , while DMSO is the solubilizing excipient in several drug products . Likewise, PEG3350 is an FDA-approved laxative (https://www.drugs.com/pro/polyethylene-glycol.html). PEGylation is used in several therapeutics to improve their pharmacokinetic and pharmacodynamic properties (https://www.biochempeg.com/article/58.html). PEG400, in particular, is an inactive ingredient in 64 FDA-approved drug formulations .DMSO and PEGs are currently part of drug formulations approved by the Food and Drug Administration (FDA). A 50% (wt/wt) DMSO solution is available for the symptomatic relief of interstitial cystitis (products , 27 (httoperties  (https:/The toxicology of these solvents has been studied extensively, with no particular concerns , 30. Non